Nanobody-enabled monitoring of kappa opioid receptor states

Tao Che (), Justin English, Brian E. Krumm, Kuglae Kim, Els Pardon, Reid H. J. Olsen, Sheng Wang, Shicheng Zhang, Jeffrey F. Diberto, Noah Sciaky, F. Ivy Carroll, Jan Steyaert, Daniel Wacker and Bryan L. Roth ()
Additional contact information
Tao Che: University of North Carolina at Chapel Hill School of Medicine
Justin English: University of North Carolina at Chapel Hill School of Medicine
Brian E. Krumm: University of North Carolina at Chapel Hill School of Medicine
Kuglae Kim: University of North Carolina at Chapel Hill School of Medicine
Els Pardon: Vrije Universiteit Brussel (VUB)
Reid H. J. Olsen: University of North Carolina at Chapel Hill School of Medicine
Sheng Wang: University of North Carolina at Chapel Hill School of Medicine
Shicheng Zhang: University of North Carolina at Chapel Hill School of Medicine
Jeffrey F. Diberto: University of North Carolina at Chapel Hill School of Medicine
Noah Sciaky: University of North Carolina at Chapel Hill School of Medicine
F. Ivy Carroll: Research Triangle Institute, Research Triangle Park
Jan Steyaert: Vrije Universiteit Brussel (VUB)
Daniel Wacker: University of North Carolina at Chapel Hill School of Medicine
Bryan L. Roth: University of North Carolina at Chapel Hill School of Medicine

Nature Communications, 2020, vol. 11, issue 1, 1-12

Abstract: Abstract Recent studies show that GPCRs rapidly interconvert between multiple states although our ability to interrogate, monitor and visualize them is limited by a relative lack of suitable tools. We previously reported two nanobodies (Nb39 and Nb6) that stabilize distinct ligand- and efficacy-delimited conformations of the kappa opioid receptor. Here, we demonstrate via X-ray crystallography a nanobody-targeted allosteric binding site by which Nb6 stabilizes a ligand-dependent inactive state. As Nb39 stabilizes an active-like state, we show how these two state-dependent nanobodies can provide real-time reporting of ligand stabilized states in cells in situ. Significantly, we demonstrate that chimeric GPCRs can be created with engineered nanobody binding sites to report ligand-stabilized states. Our results provide both insights regarding potential mechanisms for allosterically modulating KOR with nanobodies and a tool for reporting the real-time, in situ dynamic range of GPCR activity.

Date: 2020
References: Add references at CitEc
Citations: View citations in EconPapers (4)

Downloads: (external link)
https://www.nature.com/articles/s41467-020-14889-7 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-14889-7

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-020-14889-7

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().